Ogata, A. | Search bioRxiv

bioRxiv2024

CandyCollect: An Open-Microfluidic Device for the Direct Capture and Enumeration of Salivary-Extracellular Vesicles

Extracellular vesicles (EVs) are promising biomarkers for disease detection using a liquid biopsy approach, in which they are enriched and analyzed directly from biofluids. However, implementing EV biomarker technologies in the clinic remains limited by the need for practical and patient-centric biofluid collection methods that are compatible with downstream EV processing and analysis. While saliva offers a non-invasive source of EVs, its complexity and heterogeneity--cells, debris, and other non-EV proteins--can present hurdles when using traditional analytical platforms. Here, we present the CandyCollect, a lollipop-inspired sampling device with open microfluidic channels, as a patient-friendly approach for rapid salivary EV capture. CandyCollect simplifies sample preparation by effectively pre-concentrating EVs in oxygen-plasma treated open microfluidic channels. In this proof-of-principle study, we show that following a 3-5 minute-oral sampling period, EVs collected by the CandyCollect can be released with high purity within minutes and subsequently quantified and analyzed for cargo content. We observed consistent EV capture across repeated collections within individuals and expected variability across healthy participants. Additionally, single and pooled collections of EVs from a healthy participant resulted in a concordant protein profile. Overall, the CandyCollect is a new platform for rapid, non-invasive salivary EV collection and analysis for clinical diagnostics.

bioengineering↗

bioRxiv2022

Microaerophilic activated sludge system for ammonia recovery from high-strength nitrogenous wastewater: Performance and microbial communities

A transition to ammonia recovery from wastewater has started; however, a technology for sustainable nitrogen retention in the form of ammonia is still in development. This study validated a microaerophilic activated sludge (MAS) system to efficiently retain ammonia from high-strength nitrogenous wastewater. The MAS is based on conventional activated sludge (CAS) with aerobic and settling compartments. Low dissolved oxygen (DO) concentrations (<0.1 mg/L) and short solid retention times (SRTs) (<5 d) eliminated nitrifying bacteria. The two parallel MASs were successfully operated for 300 d and had ammonia retention of 101.7 {+/-} 24.9% and organic carbon removal of 85.5 {+/-} 8.9%. The MASs mitigated N2O emissions with an emission factor of <0.23%, much lower than the default value of CAS (1.6%). A short-term step-change test demonstrated that N2O indicated the initiation of nitrification and the completion of denitrification in the MAS. The parallel MASs had comparable microbial diversity, promoting organic carbon oxidation while inhibiting ammonia-oxidizing microorganisms (AOMs), as revealed by 16S rRNA gene amplicon sequencing, qPCR of functional genes, and fluorescent in situ hybridization of {beta}-Proteobacteria AOB. The microbial analyses also uncovered that filamentous bacteria were positively correlated with effluent turbidity. Together, controlling DO and SRT achieved successful ammonia retention, mainly by suppressing AOM activity. This process represents a new nitrogen management paradigm. SynopsisMoving from nitrogen removal to nitrogen recovery is critical for establishing a sustainable society. We provided proof-of-the-concept for a novel ammonia retention technology by retrofitting an activated sludge system.

microbiology↗